Daniel:Notebook/ComboLock/2017-4-12: Difference between revisions

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>Djacobse
>Djacobse
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{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt;font-weight:bold" align="center"
|- style="font-size:12pt;font-weight:bold" align="center"
|style="background-color:#8DB4E2" width="250" height="30" | Reagent
|style="background-color:#8DB4E2" width="230" height="30" | Reagent
|style="background-color:#8DB4E2" width="85" | Single Rxn Vol
|style="background-color:#8DB4E2" width="85" | Single Rxn Vol
|style="background-color:#95B3D7" width="85" | Master Mix (13.2X)
|style="background-color:#95B3D7" width="85" | Master Mix (13.2X)
Line 18: Line 18:
|- style="font-size:12pt"
|- style="font-size:12pt"
| height="15"  valign="bottom" | RCA Rxn
| height="15"  valign="bottom" | RCA Rxn
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 7
| align="center" align="center" valign="bottom" | 0
| align="center" align="center" valign="bottom" | 0


|- style="background-color:#BFBFBF;font-size:12pt"
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | Bglii Cutter Primer + (100 uM)
| height="15"  valign="bottom" | 10X Buffer 3.1
| align="center" align="center" valign="bottom" | 2
| align="center" align="center" valign="bottom" | 2
| align="center" align="center" valign="bottom" | 26.4
| align="center" align="center" valign="bottom" | 26.4


|- style="font-size:12pt"
|- style="font-size:12pt"
| height="15"  valign="bottom" | 10X Buffer 3.1
| align="center" align="center" valign="bottom" | 2
| align="center" align="center" valign="bottom" | 26.4
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | Bglii
| height="15"  valign="bottom" | Bglii
| align="center" align="center" valign="bottom" | 1
| align="center" align="center" valign="bottom" | 1
| align="center" align="center" valign="bottom" | 0
| align="center" align="center" valign="bottom" | 0


|- style="font-size:12pt"
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | nfH2O
| height="15"  valign="bottom" | nfH2O
| align="center" align="center" valign="bottom" | 10
| align="center" align="center" valign="bottom" | 10
| align="center" align="center" valign="bottom" | 132
| align="center" align="center" valign="bottom" | 132


|- style="background-color:#BFBFBF;font-size:12pt"
|- style="font-size:12pt"
| height="15"  valign="bottom" | Total
| height="15"  valign="bottom" | Total
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20
|style="font-weight:bold" align="center" align="center" valign="bottom" | 184.8
|style="font-weight:bold" align="center" align="center" valign="bottom" | 158.4


|}
|}


<li>Aliquot 14 uL master mix into new tube</li>
<li>Aliquot 12 uL master mix into new tube</li>
<li>Add 5 uL appropriate sample</li>
<li>Add 7 uL appropriate sample</li>
<li>Use the following thermocycler program</li>
<li>Use the following thermocycler program</li>
<ol type="a">
<ol type="a">

Revision as of 15:13, 12 April 2017

RCA Double Deuce

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Protocol

  1. Bglii digestion
    1. Make the following reactions; Do not add Bglii yet
    2. Reagent Single Rxn Vol Master Mix (13.2X)
      RCA Rxn 7 0
      10X Buffer 3.1 2 26.4
      Bglii 1 0
      nfH2O 10 132
      Total 20 158.4
    3. Aliquot 12 uL master mix into new tube
    4. Add 7 uL appropriate sample
    5. Use the following thermocycler program
      1. 2 hr 37C; When this cycle starts add 1 uL Bglii
      2. Heat kill with 65C for 20 min